CAL1 is the Drosophila CENP-A assembly factor

J Cell Biol. 2014 Feb 3;204(3):313-29. doi: 10.1083/jcb.201305036. Epub 2014 Jan 27.

Abstract

Centromeres are specified epigenetically by the incorporation of the histone H3 variant CENP-A. In humans, amphibians, and fungi, CENP-A is deposited at centromeres by the HJURP/Scm3 family of assembly factors, but homologues of these chaperones are absent from a number of major eukaryotic lineages such as insects, fish, nematodes, and plants. In Drosophila, centromeric deposition of CENP-A requires the fly-specific protein CAL1. Here, we show that targeting CAL1 to noncentromeric DNA in Drosophila cells is sufficient to heritably recruit CENP-A, kinetochore proteins, and microtubule attachments. CAL1 selectively interacts with CENP-A and is sufficient to assemble CENP-A nucleosomes that display properties consistent with left-handed octamers. The CENP-A assembly activity of CAL1 resides within an N-terminal domain, whereas the C terminus mediates centromere recognition through an interaction with CENP-C. Collectively, this work identifies the "missing" CENP-A chaperone in flies, revealing fundamental conservation between insect and vertebrate centromere-specification mechanisms.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoantigens / metabolism*
  • Cell Line
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Chromosome Segregation
  • DNA, Superhelical / metabolism
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / metabolism*
  • Epigenesis, Genetic
  • Histones / metabolism
  • Kinetochores / metabolism
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism
  • Nucleosomes / metabolism
  • Protein Binding
  • Protein Sorting Signals

Substances

  • Autoantigens
  • CENPA protein, human
  • Cal1 protein, Drosophila
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • DNA, Superhelical
  • Drosophila Proteins
  • Histones
  • Microtubule-Associated Proteins
  • Ndc80 protein, Drosophila
  • Nucleosomes
  • Protein Sorting Signals
  • centromere protein C